Collection and Culture of Human Connective Tissue Cells from Gingival Explant Technique for Oral Tissue Bioengineering

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Fernando Antônio Mauad de Abreu; Igor Daniel Garcia Reis; Gerluza Aparecida Borges Silva & Erika Cristina Jorge


Cell culture is an important tool in medical, odontological and biological research laboratories, supporting cell therapies and tissue bioengineering strategies. Gingival fibroblasts present structural function, being able to modulate their metabolic capacity, which is reflected in the tissue morphology. The possibility of culturing fibroblasts in vitro, in monolayer or on three-dimensional scaffolds, for subsequent transplants in vivo opens important perspectives for the periodontal surgical clinic. The objective of the present article is to present a method of obtaining and cultivating viable human gingival fibroblasts for in vitro research. Explants derived from periodontal surgical discards were used, grown in 25 cm2 bottles to obtain a primary cell culture. After observing the proliferation and growth of the fibroblasts that interconnected and formed a monolayer network, involving the periphery of the explants, it was possible to remove the explants, to make the passage and the new subcultures were obtained in a ratio of 1:1. After 7 days, the amount of viable cells was analyzed in triplicate, using the Neubauer chamber technique, in cell culture bottles of 25 mm2 (T25) and 75 mm2 (T75). Fibroblasts were described and subclassified morphologically. The results showed a growth pattern in both bottles, but with a larger number in bottles of 75 cm2. Cells with fibroblastic morphology were subclassified into reticular and fusiform, being predominant those with fusiform morphology. In conclusion, culture of explant of human gingival connective tissue is a viable method for obtaining gingival connective tissue cells suitable for laboratory tests in cell culture, aiming at obtaining constructs for gingival tissue engineering.

KEY WORDS: Cell culture technique; Primary cell culture; Connective tissue Cells; Cellular biology.

How to cite this article

ABREU, F. A. M.; REIS, I. D. G.; SILVA, G. A. B. & JORGE, E. C. Collection and culture of human connective tissue cells from gingival explant technique for oral tissue bioengineering. Int. J. Morphol., 37(4):1229-1233, 2019.